Abstract:

BACKGROUND: Streptococcus mutans is the main pathogenic bacterium for dental caries. In terms of the gene vaccine's SpaP which mediate Streptococcus mutans non-sucrose dependent adherency, it antagonizes academically Streptococcus mutans, which adheres to dental facing and makes the tooth tissue worse.
OBJECTIVE: To construct eukanyotic plasmid pVAX1-spap/P of surface protein antigen P of Streptococcus mutans and to evaluate the expression of the plasmid in mammalian cells COS-7.
METHODS: The eukaryotic plasmid carrying encoding gene of spap/P of Streptococcus mutans was constructed and then analyzed by the restriction enzyme and sequence correctly, the plasmid was introduced into COS-7 cells by lipofectamine reagent. The transient expressed protein was detected by immunochemistry technique in COS-7 cells.
RESULTS AND CONCLUSION: The expression of recombinant plasmid pVAX1-spap/P analyzed by the EcoR Ⅰ and Xba Ⅰrestriction enzyme confirmed carrying spap/P fragment of 1.2 kb. After sequencing, target gene was positively inserted into pre-designed point of the vector spaces. Positive expression was detected in plasma of the cells which were transfected with recombinant plasmid pVAX1-spap/P. The cells which were transfected with pVAX1 were negative. The eukaryotic plasmid pVAX1-spap/P was constructed successfully and target gene spap/P translated and expressed in COS-7 cells after transfecting with recombinant plasmid pVAX1-spap/P.